IEF pattern classification-derived criteria for the identification of epoetin-delta in urine

Epoetin-δ (Dynepo™ Shire Pharmaceuticals, Basing stoke, UK) is a synthetic form of erythropoietin (EPO) whose resemblance with endogenous EPO makes it hard to identify using the classical identification criteria. Urine samples collected from six healthy volunteers treated with epoetin-δ injections and from a control population were immuno-purified and analyzed with the usual IEF method. On the basis of the EPO profiles integration, a linear multivariate model was computed for discriminant analysis. For each sample, a pattern classification algorithm returned a bands distribution and intensity score (bands intensity score) saying how representative this sample is of one of the two classes, positive or negative. Effort profiles were also integrated in the model. The method yielded a good sensitivity versus specificity relation and was used to determine the detection window of the molecule following multiple injections. The bands intensity score, which can be generalized to epoetin-α and epoetin-β, is proposed as an alternative criterion and a supplementary evidence for the identification of EPO abuse.

Response to long-distance relocation in Asian elephants (Elephas maximus) : monitoring adrenocortical activity via serum, urine, and feces

Understanding how elephants respond to potentially stressful events, such as relocation, is important for making informed management decisions. This study followed the relocation of eight Asian elephants from the Cocos (Keeling) Islands to mainland Australia. The first goal of this study was to examine patterns of adrenocortical activity as reflected in three different substrates: serum, urine, and feces. We found that the three substrates yielded very different signals of adrenocortical activity. Fecal glucocorticoid metabolites (FGM) increased as predicted post-transport, but urinary glucocorticoid metabolites (UGM) were actually lower following transport. Serum cortisol levels did not change significantly. We suggest that the differences in FGM and UGM may reflect changes in steroid biosynthesis, resulting in different primary glucocorticoids being produced at different stages of the stress response. Additional studies are needed to more thoroughly understand the signals of adrenocortical activity yielded by different substrates. The second goal was to examine individual variation in patterns of adrenal response. There was a positive correlation between baseline FGM value and duration of post-transfer increase in FGM concentration. Furthermore, an individual's adrenocortical response to relocation was correlated with behavioral traits of elephants. Elephants that were described by keepers as being “curious” exhibited a more prolonged increase in FGM post-transfer, and “reclusive” elephants had a greater increase in FGM values. Future research should investigate the importance of these personality types for the management and welfare of elephants.

Grain excretion by goats fed whole or processed cereals with various roughages

Despite cereal grains being grown on 5 continents where goats are kept, there is little information on the excretion of whole cereal grains when fed to goats. We determined the effects of various dietary treatments on whole grain and starch loss in the faeces of Angora goats. In Experiment 1 there were 4 replicates of the factorial design: (a) 2 grain types (barley, oats); (b) whole grain or processing (milled barley or rolled oats); (c) 2 roughage qualities (Persian clover hay, barley straw); and (d) 2 feeding levels (level 1, 150 g/d of grain, 250 g/d of roughage; level 2, 250 g/d of grain, ad libitum roughage). In Experiment 2, which immediately followed Experiment 1, and aimed to detect carry over effects of previous feeding of barley straw and grain processing, feed levels were either 650 g/d grain or 400 g/d grain with 550 g/d Persian clover hay. Data were analysed by ANOVA. In Experiment 1, processing had no effect on digestible dry matter intake. The number of whole grains lost per 100 g of fresh faeces and whole grains loss as the % of grain dry matter intake were affected by an interaction between processing and roughage quality. Whole grain fed with Persian clover hay had greater grain loss than all other diets. Whole grain loss was greater with whole grain than with processed grain. Level of feeding had no effects on grain loss. In Experiment 2, more whole grains were lost in fresh faeces when fed with Persian clover hay than when fed without hay, an effect of previous feeding with barley straw reduced whole grain excretion, and more barley grains were lost than oat grain. Faecal starch was affected, with higher levels when whole barley grain was fed, particularly with Persian clover hay, or when previously fed barley straw at a high level. Feeding grain at 650 g/d did not increase grain or starch excretion. Whole grains represented a small loss of grain dry matter intake in faeces, averaging 0.8% with a maximum recorded of 2.6%. Faecal concentration of the whole grains may be altered by grain size and the digestibility of the roughage component of the diet. In this study an additional cost of 3% for processing grains would not have provided economic benefits.

Direct processing of clinically relevant large volume samples for the detection of sexually transmitted infectious agents from urine on a microfluidic device

Urine is a preferred specimen for nucleic acid-based detection of sexually transmitted infections (STIs) but represents a challenge for microfluidic devices due to low analyte concentrations. We present an extraction methodology enabling rapid on-chip nucleic acid purification directly from clinically relevant sample volumes up to 1 ml and subsequent PCR amplification detection.

Altered responsiveness of the kidney to activation of the renal nerves in fat-fed rabbits

We tested whether mild adiposity alters responsiveness of the kidney to activation of the renal sympathetic nerves. After rabbits were fed a high-fat or control diet for 9 wk, responses to reflex activation of renal sympathetic nerve activity (RSNA) with hypoxia and electrical stimulation of the renal nerves (RNS) were examined under pentobarbital anesthesia. Fat pad mass and body weight were, respectively, 74% and 6% greater in fat-fed rabbits than controls. RNS produced frequency-dependent reductions in renal blood flow, cortical and medullary perfusion, glomerular filtration rate, urine flow, and sodium excretion and increased renal plasma renin activity (PRA) overflow. Responses of sodium excretion and medullary perfusion were significantly enhanced by fat feeding. For example, 1 Hz RNS reduced sodium excretion by 79 ± 4% in fat-fed rabbits and 46 ± 13% in controls. RNS (2 Hz) reduced medullary perfusion by 38 ± 11% in fat-fed rabbits and 9 ± 4% in controls. Hypoxia doubled RSNA, increased renal PRA overflow and medullary perfusion, and reduced urine flow and sodium excretion, without significantly altering mean arterial pressure (MAP) or cortical perfusion. These effects were indistinguishable in fat-fed and control rabbits. Neither MAP nor PRA were significantly greater in conscious fat-fed than control rabbits. These observations suggest that mild excess adiposity can augment the antinatriuretic response to renal nerve activation by RNS, possibly through altered neural control of medullary perfusion. Thus, sodium retention in obesity might be driven not only by increased RSNA, but also by increased responsiveness of the kidney to RSNA.

The association of knowledge, attitudes and behaviours related to salt with 24-hour urinary sodium excretion

Salt reduction efforts usually have a strong focus on consumer education. Understanding the association between salt consumption levels and knowledge, attitudes and behaviours towards salt should provide insight into the likely effectiveness of education-based programs.

Are urinary porphyrins a valid diagnostic biomarker of autism spectrum disorder?

A fundamental challenge to the timely diagnosis of Autism Spectrum Disorder (ASD) is the reliance on the observation of a set of aberrant behavior. Consequently, the diagnostic process requires that the child reach an age where the behaviors would typically be exhibited. The identification of a reliable biological marker (biomarker) could be of considerable benefit to the diagnostic process. As a diagnostic biomarker, porphyrins present an attractive prospect as previous studies have reported consistent findings of children with ASD showing significant elevations in porphyrin levels in contrast to controls. Furthermore, there is some evidence that ASD severity may be associated with porphyrins, which would be a valuable characteristic of any ASD biomarker. Importantly, for practical use, porphyrins can be tested non-invasively via a sample of urine. The present study sought to investigate whether porphyrin profiles can reliably be used to (a) differentiate ASD cases from healthy controls; and (b) predict ASD severity. The study compared the porphyrin levels of three groups of children aged 2-6 years: Group 1-children diagnosed with ASD (n = 70); Group 2-healthy, normally developing siblings of children diagnosed with ASD (n = 36); and Group 3-healthy, normally developing children with no known blood relative diagnosed with ASD (n = 54). The results of logistic regression analyses failed to find support for the hypotheses that porphyrin levels could be used as a valid tool to detect ASD cases or predict severity. Autism Res 2014, 7: 535-542. © 2014 International Society for Autism Research, Wiley Periodicals, Inc.

Self-monitoring of blood glucose versus self-monitoring of urine glucose in adults with newly diagnosed Type 2 diabetes receiving structured education: a cluster randomized controlled trial.

AIMS: To compare the effectiveness and acceptability of self-monitoring of blood glucose with self-monitoring of urine glucose in adults with newly diagnosed Type 2 diabetes. METHODS: We conducted a multi-site cluster randomized controlled trial with practice-level randomization. Participants attended a structured group education programme, which included a module on self-monitoring using blood glucose or urine glucose monitoring. HbA1c and other biomedical measures as well as psychosocial data were collected at 6, 12 and 18 months. A total of 292 participants with Type 2 diabetes were recruited from 75 practices. RESULTS: HbA1c levels were significantly lower at 18 months than at baseline in both the blood monitoring group [mean (se) -12 (2) mmol/mol; -1.1 (0.2) %] and the urine monitoring group [mean (se) -13 (2) mmol/mol; -1.2 (0.2)%], with no difference between groups [mean difference adjusted for cluster effect and baseline value = -1 mmol/mol (95% CI -3, 2); -0.1% (95% CI -0.3, 0.2)]. Similar improvements were observed for the other biomedical outcomes, with no differences between groups. Both groups showed improvements in total treatment satisfaction, generic well-being, and diabetes-specific well-being, and had a less threatening view of diabetes, with no differences between groups at 18 months. Approximately one in five participants in the urine monitoring arm switched to blood monitoring, while those in the blood monitoring arm rarely switched (18 vs 1% at 18 months; P < 0.001). CONCLUSIONS: Participants with newly diagnosed Type 2 diabetes who attended structured education showed similar improvements in HbA1c levels at 18 months, regardless of whether they were assigned to blood or urine self-monitoring.

Dietary salt intake and discretionary salt use in two general population samples in Australia: 2011 and 2014

The limited Australian measures to reduce population sodium intake through national initiatives targeting sodium in the food supply have not been evaluated. The aim was, thus, to assess if there has been a change in salt intake and discretionary salt use between 2011 and 2014 in the state of Victoria, Australia. Adults drawn from a population sample provided 24 h urine collections and reported discretionary salt use in 2011 and 2014. The final sample included 307 subjects who participated in both surveys, 291 who participated in 2011 only, and 135 subjects who participated in 2014 only. Analysis included adjustment for age, gender, metropolitan area, weekend collection and participation in both surveys, where appropriate. In 2011, 598 participants: 53% female, age 57.1(12.0)(SD) years and in 2014, 442 participants: 53% female, age 61.2(10.7) years provided valid urine collections, with no difference in the mean urinary salt excretion between 2011: 7.9 (7.6, 8.2) (95% CI) g/salt/day and 2014: 7.8 (7.5, 8.1) g/salt/day (p = 0.589), and no difference in discretionary salt use: 35% (2011) and 36% (2014) reported adding salt sometimes or often/always at the table (p = 0.76). Those that sometimes or often/always added salt at the table and when cooking had 0.7 (0.7, 0.8) g/salt/day (p = 0.0016) higher salt excretion. There is no indication over this 3-year period that national salt reduction initiatives targeting the food supply have resulted in a population reduction in salt intake. More concerted efforts are required to reduce the salt content of manufactured foods, together with a consumer education campaign targeting the use of discretionary salt.

Cross-platform urine metabolomics of experimental hyperglycemia in type 2 diabetes

Hyperglycemia causes diabetic nephropathy, a condition for which there are no specific diagnostic markers thatpredict progression to renal failure. Here we describe a multiplatform metabolomic analysis of urine from individualswith type 2 diabetes, collected before and immediately following experimental hyperglycemia. We used targetednuclear magnetic resonance spectroscopy (NMR), liquid chromatography - mass spectrometry (LC-MS) and gaschromatography - MS (GC-MS) to identify markers of hyperglycemia. Following optimization of data normalisation andstatistical analysis, we identified a reproducible NMR and LC-MS based urine signature of hyperglycemia. Significantincreases of alanine, alloisoleucine, isoleucine, leucine, N-isovaleroylglycine, valine, choline, lactate and taurine anddecreases of arginine, gamma-aminobutyric acid, hippurate, suberate and N-acetylglutamate were observed. GC-MSanalysis identified a number of metabolites differentially present in post-glucose versus baseline urine, but these could not be identified using current metabolite libraries. This analysis is an important first step towards identifying biomarkers of early-stage diabetic nephropathy.

Estudo clínico e bioquímico de 28 pacientes com mucopolissacaridose tipo VI

Realizamos um estudo observacional de pacientes com mucopolissacaridose tipo VI, com o objetivo de determinar o perfil epidemiológico, clínico e bioquímico de um grupo de pacientes sul-americanos a fim de contribuir em estudos futuros de correlação genótipo-fenótipo e de avaliação de protocolos clínicos. Os critérios de inclusão foram: ter 4 anos ou mais e confirmação bioquímica da doença (níveis reduzidos da atividade da ARSB, aumento de GAGs urinários e atividade normal de outra sulfatase). Os critérios de exclusão foram: terapia com reposição enzimática atual ou prévia ou ter realizado transplante de medula óssea. Foram avaliados 28 pacientes por anamnese, exame físico, acocardiograma, eletrocardiograma, avaliação oftalmológica, medidas de glicosaminoglicanos urinários e da atividade da N-acetilgalactosamina-4-sulfatase em leucócitos. A amostra estudada tinha 92,9% de brasileiros, sendo 53,8% da região sudeste. No momento da avaliação, a média de idade foi de 97,1 meses e a média de idade ao diagnóstico foram de 48,4 meses. Em 88% da amostra os sintomas iniciaram com menos de 36 meses e em 27% das famílias houve relato de consangüinidade entre os pais. A média de peso e estatura ao nascimento foi de 3481 gramas e 51,3 centímetros, respectivamente. Da amostra, 57,1% nasceram de parto vaginal. Todos apresentavam alguma alteração ecocardiográfica, bem como opacificação corneana. As manifestações clínicas mais freqüentes foram: baixa estatura, opacificação corneana, facies grosseira, contraturas articulares e mãos em garra. A média da atividade enzimática em leucócitos foi de 5,4 nmoles/h x mg proteína e a excreção urinária de glicosaminoglicanos foi, em média, 7,9 vezes superior ao normal. O número de manifestações clínicas citadas não apresentou correlação significativa com a idade, com a excreção urinária de GAGs ou com a atividade enzimática em leucócitos. Também não houve correlação significativa entre a excreção urinária de GAGS e a atividade enzimática. Concluímos que a MPS VI é uma patologia com alta morbidade e que, comparados com a literatura, os pacientes da nossa amostra têm um diagnóstico tardio e maior freqüência de alterações cardiológicas.